Dried biological compositions and methods thereof

Abstract

Dried and stable biological compositions may have high colony-forming units. Such compositions may include (1) a substrate and (2) micro-organism(s) loaded onto the surface of the substrate, wherein the composition has a total moisture content of about 0.01 wt. % to about 15 wt. %, preferably, about 0.01 wt. % to about 8 wt. %, preferably, about 3 wt. % to about 8 wt. %, preferably, about 5 wt. % to about 8 wt. %, still preferably, selected from 3 wt. %, 5 wt. % and 7 wt. %.

1 . A dried biological composition, comprising:
 (1) a substrate; and   (2) micro-organisms loaded onto the surface of the substrate,   wherein the composition has a total moisture content in a range of from 0.01 to 15 wt. %.   
     
     
         2 . The composition of  claim 1 , wherein the composition has a water activity value (Aw) in a range of from 0.01 to 0.6. 
     
     
         3 . The composition of  claim 1 , wherein the composition has greater than 107 CFU/g. 
     
     
         4 . The composition of  claim 1 , wherein the substrate is selected from the group consisting of silica, diatomaceous earth, silica gel, silicates, and water-insoluble natural fiber material. 
     
     
         5 . (canceled) 
     
     
         6 . The composition of  claim 1 , wherein the substrate is precipitated silica. 
     
     
         7 . (canceled) 
     
     
         8 . The composition of  claim 1 , wherein a particle size (d50) of the substrate is in a range of from 5 to 200 microns. 
     
     
         9 . The composition of  claim 1 , wherein a BET surface area of the substrate is in a range of from 2 to 600 m 2 /g. 
     
     
         10 - 11 . (canceled) 
     
     
         12 . The composition of  claim 1 , wherein a pore volume of the substrate is in a range of from 0.01 to 1.20 cc/g, based on the Barrett-Joyner-Halenda model, or
 wherein a BET surface area of the substrate is in a range of from 400 to 600 m 2 /g, and with a pore volume of greater than 1 cc/g, by Barrett-Joyner-Halenda model or greater than 2 cc/g by Mercury Pore Volume.   
     
     
         13 . The composition of  claim 1 , wherein a final microorganism concentration is in a range of from 4 to 40 wt. %, based on total composition weight. 
     
     
         14 . The composition of  claim 1 , wherein the micro-organisms are selected from the group consisting of  Bacillus subtilis  QST713 , Pasteuria  usgae;  Beauveria bassiana, Coniothyrium minitans, Chondrostereum purpureum, Paecilomyces lilacinus, Aschersonia aleyrodis, Beauveria brongniartii, Hirsutella thompsonii, Isaria fumosorosea, Isaria  sp.,  Lecanicillium longisporum, Lecanicillium muscarium, Lecanicillium  sp.,  Metarhizium anisopliae, Metarhizium anisopliae  var.  acridum, Nomuraea rileyi Sporothrix insectorum; Cydia pomonella  GV;  Phytophthora palmivora, Lagenidium giganteum, Bacillus thuringiensis, Pseudomonas fluorescens, Bradyrhizobium, Mycorrhiza, Clonostachys rosea, Bacillus  spp. and  Lactobacillus  spp., and a combination thereof. 
     
     
         15 . The composition of  claim 1 , wherein the micro-organisms are  Clonostachys rosea  or  Pseudomonas fluorescens.    
     
     
         16 . The composition of  claim 1 , in tablet form, flowable concentrate form, or in oil dispersion form further comprising one or more excipient. 
     
     
         17 . The composition of  claim 1 , not requiring exogenous protectant. 
     
     
         18 . The composition of  claim 1 , further comprising:
 (i) a polymer selected from the group consisting of polyvinyl alcohol, xanthan gum, gum arabic, a different polysaccharide, polyethylene glycol, and polyglycerol; or   (ii) non-reducing disaccharide; or   (iii) skim milk or dimethyl sulfoxide.   
     
     
         19 . The composition of  claim 1 , further comprising:
 a second substrate as an outer layer, preferably, the second substrate is selected from (i) a precipitated silica and (ii) a fumed silica.   
     
     
         20 . The composition of  claim 1 , wherein a number of colony forming units per gram of the composition (CFU/g) remains above 107 CFU/g after storage at:
 (a) room temperature for 120 days;   (b) 40° C. for 40 days; and/or   (c) a relative humidity of 65% or lower for 40 days.   
     
     
         21 . The composition of  claim 1 , having a tap density greater than 150% of a tapped density of a pure substrate material. 
     
     
         22 . A process for preparing a dried biological composition, the method comprising:
 (1) combining a micro-organism mixture, solution, or suspension with a substrate, to obtain a substrate-microorganism mixture; and   (2) drying the substrate-microorganism mixture to reach a total moisture content in a range of from 0.01 to 15 wt. %.   
     
     
         23 . The process of  claim 22 , wherein the microorganisms are harvested from a surface of a seed by (a) mechanically grinding or polishing the surface of the substrate resulting in a fine fraction including microorganisms and some parts of the seed; and optionally (b) sieving the resulting fine fraction in order to obtain a powder with a defined particle size distribution for subsequent processing. 
     
     
         24 . The process of  claim 23 , wherein the mechanically grinding or polishing (a) comprises grinding the seed with a grinding stone to separate the seed from the fine fraction 
     
     
         25 . The process of  claim 23 , wherein the mechanically grinding or polishing (a) comprises grinding with a rotating shaft inside an enclosing tube of a slotted screen at an under pressure, followed by contacting with a sifter and a filter to separate the seed form the fine fraction. 
     
     
         26 . The process of  claim 23 , wherein the sieving (b) of the fine fraction is conducted and comprises sieving with a sieve mesh size in a range of from 20 to 800 μm. 
     
     
         27 . The process of  claim 22 , wherein the microorganisms are harvested from a surface of a seed by washing them off with water and separating the seed and a liquid microorganism solution or suspension. 
     
     
         28 . The process of  claim 22 , wherein drying (2) comprises fluid bed drying, spray drying, contact drying, or freeze drying the substrate-microorganism mixture. 
     
     
         29 - 31 . (canceled) 
     
     
         32 . The process of  claim 22 , wherein a drying air temperature is less than or equal to 130° C. 
     
     
         33 . The process of  claim 22 , wherein a powder bed is maintained at less than or equal to 35° C. 
     
     
         34 . The process of  claim 22 , wherein the resulting composition has a water activity value (Aw) in a range of from 0.01 to 0.6. 
     
     
         35 . The process of  claim 22 , wherein a resulting composition from the drying has a colony-forming units of microorganisms per gram of composition greater than 107 CFU/g. 
     
     
         36 . The process of  claim 22 , wherein the substrate is selected from the group consisting of silica, diatomaceous earth, silica gel, silicates, and water-insoluble natural fiber material. 
     
     
         37 . The process of  claim 22 , wherein the substrate is silica. 
     
     
         38 . The process of  claim 22 , wherein the substrate has a BET surface area in a range of from 400 to 600 m 2 /g by Barrett-Joyner-Halenda model, and the pore volume of great than 1 cc/g by Mercury Pore Volume. 
     
     
         39 . The process of  claim 22 , wherein the substrate is selected from those having:
 (i) a particle size (d50) in a range of from 5 to 200 microns;   (ii) a BET surface area in a range of from 2 to 400 m 2 /g;   (iii) a pore volume of the substrate is in a range of from 0.01 to 1.20 cc/g;   (iv) or any combination thereof.   
     
     
         40 . The process of  claim 22 , wherein the combining (1) comprises loading an amount in a range of from 4 to 40 wt. %, of the total composition. 
     
     
         41 . The process of  claim 22 , wherein micro-organisms are selected from the group consisting of  Bacillus subtilis  QST713 , Pasteuria usgae; Beauveria bassiana, Coniothyrium minions, Chondrostereum purpureum, Paecilomyces lilacinus, Aschersonia aleyrodis, Beauveria brongniartii, Hirsutella thompsonii, Isaria fumosorosea, Isaria  sp.,  Lecanicillium longisporum, Lecanicillium muscarium, Lecanicillium  sp.,  Metarhizium anisopliae, Metarhizium anisopliae  var.  acridum, Nomuraea rileyi Sporothrix insectorum; Cydia pomonella  GV;  Phytophthora palmivora, Lagenidium giganteum, Bacillus thuringiensis, Pseudomonas fluorescens, Bradyrhizobium, Mycorrhiza, Clonostachys rosea, Bacillus  spp.,  Lactobacillus  spp., and a combination thereof. 
     
     
         42 . The process of  claim 22 , wherein the resulting composition does not require exogenous protectant. 
     
     
         43 . A dry biological composition, prepared by the process of  claim 22 . 
     
     
         44 . The composition  claim 1 , further comprising:
 a seed to be treated.   
     
     
         45 . A method for controlling insect, fungus, or nematode on an area to be treated, the method comprising:
 optionally reconstituting the dry biological composition of  claim 1 , and   applying an effective amount of the optionally reconstituted composition to the area to affect treatment.   
     
     
         46 . The method of  claim 45 , wherein the area to be treated is a portion of a plant.